Gene/Protein Disease Symptom Drug Enzyme Compound
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Query: EC:3.1.30.2 (endonuclease)
 18,621 document(s) hit in 31,850,051 MEDLINE articles (0.00 seconds)

To analyze the characteristics of Leptospira strains HY-1, HY-2, and HY-10, which were isolated from patients with leptospirosis in Korea in 1985, 12 monoclonal antibodies (MAbs) against strain HY-1 and six MAbs against serovar lai strain 017 were produced, and their properties were determined by the microscopic agglutination test. Genetic relationships among the leptospires were determined by restriction endonuclease DNA analysis. Three MAbs reacted with all strains of serogroup Icterohaemorrhagiae, but did not react with any strains of the other 10 serogroups. All MAbs reacted with strains 017, HY-1, HY-2, and HY-10 at nearly identical titers. Two MAbs reacted only with these four strains. These four strains also had the same restriction endonuclease cleavage patterns. Based on these results, strains HY-1, HY-2, and HY-10 were identified as serovar lai, which is one of the common serovars in China. It is suggested that serovar lai is one of the prevalent serovars in Korea, and that the Mabs produced in this study are useful for the accurate and rapid identification of this serovar.
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PMID:Characterization of Leptospira strains HY-1, HY-2, and HY-10 isolated in Korea by means of monoclonal antibodies and restriction endonuclease DNA analysis. 155 73

Leptospira interrogans serovar icterohaemorrhagiae strains Ictero No. I and RGA and serovar copenhageni strains M20, Shiromizu and Shibaura were examined by restriction endonuclease DNA analysis. Fifteen endonucleases (AluI, BamHI, BglII, EcoRI, HaeIII, HhaI, HindIII, KpnI, PstI, SacI, SalI, SmaI, StyI, XbaI and XhoI) were used as the digesting enzymes. Strain Ictero No. I showed endonuclease cleavage patterns which differed from those of the other four strains only when it was digested with enzymes KpnI and HindIII. When digested with KpnI, an extra band of about 5.4 kb was clearly produced, and when digested with HindIII, an extra band of about 25 kb was produced. When the other 13 enzymes were used, no differences were found between the endonuclease cleavage patterns among the five strains. Moreover, strains RGA, M20, Shiromizu and Shibaura could not be distinguished by the restriction endonuclease DNA analysis using all 15 endonucleases. In addition, six newly isolated leptospires from patients with leptospirosis and from Rattus norvegicus were compared with the Ictero No. I and M20 strains, by restriction endonuclease DNA analysis using enzymes KpnI and HindIII. Three leptospires belonging to serovar icterohaemorrhagiae showed the same endonuclease cleavage patterns as the M20 strain. The other three strains, which belong to serovar copenhageni, showed almost the same endonuclease cleavage patterns as the M20 strain; only the Kai ima 702 strain produced an extra band which was not identical to the Ictero No. I-specific extra band when digested with HindIII. The leptospiral restriction endonuclease DNA analysis has revealed taxonomic structures that are unrecognized by serology alone.
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PMID:Restriction endonuclease DNA analysis of Leptospira interrogans serovars Icterohaemorrhagiae and Copenhageni. 285 Apr 47

The authors describe work in progress at the laboratory in Brescia, Italy, on the application of molecular methods to the diagnosis of leptospirosis. This work includes the following: a) Development of polymerase chain reaction (PCR) assays capable of amplifying specific deoxyribonucleic acid fragments from most Leptospira interrogans strains. b) Development of a microtitre-based assay for rapid detection of PCR-positive samples. c) Characterisation of Leptospira strains through restriction endonuclease analysis of PCR products and amplified fragment length polymorphism.
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PMID:The search for improved methods for diagnosing leptospirosis: the approach of a laboratory in Brescia, Italy. 810 48

Leptospira, a member of the order Spirochaetales, is the causative agent of leptospirosis, an important zoonosis encountered worldwide. The Leptospira interrogans serovar Sejroe was grown in EMJH medium and its DNA was isolated using standard techniques. The LipL32 gene was amplified using the reported primer of Kirschneri of LipL32. The amplified product was found to comprise 756 base pairs. This amplified gene fragment of LipL32 lipoprotein was cloned in E. coli (DH5 alpha) cells using pDrive plasmid as a vector. The recombinant cells were selected on LB agar medium containing ampicillin, X-gal and isopropyl-beta-D-thiogalactopyranoside. Plasmid was extracted from the recombinant white colonies, and restriction endonuclease (RE) analysis was carried out using PstI and SalI. On partial sequence analysis, the product exhibited 756 base pairs, corresponding to 251 amino acids. The cloned gene could be further used for expression of recombinant protein for serodiagnosis of leptospirosis.
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PMID:Cloning and sequence analysis of the gene encoding LipL32 of Leptospira interrogans serovar Sejroe. 1722 65

Leptospirosis is an emerging infectious disease, which is considered to be the most widespread zoonotic disease in the world. There are more than 230 known serovars in the genus Leptospira. A loop-mediated isothermal amplification (LAMP) assay for the rapid detection of pathogenic Leptospira spp. was developed and evaluated through amplification of the lipL41 gene coding for the outer membrane protein LipL41. The LAMP assay did not rely on the isolation and culture of leptospires, and no cross-reactivity was observed with other bacterial species. A SYBR Green I-based LAMP assay was also carried out for the real-time detection of DNA amplification. The lower detection limit of the LAMP assay was approximately 100 copies, which was the same as the polymerase chain reaction (PCR) and real-time PCR assays. The accuracy of the LAMP reaction was confirmed by restriction endonuclease analysis of the amplified product. The LAMP assay is easy to perform and inexpensive, and so may be applied in the rapid and specific diagnosis of Leptospira.
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PMID:Application of a loop-mediated isothermal amplification method for the detection of pathogenic Leptospira. 1907 Apr 50

The incidence of leptospirosis in human beings has been increasing in recent years. Early diagnosis and treatment can prevent complications and reduce mortality. The conventional laboratory methods for diagnosis rely on the demonstration of leptospires in clinical specimens, recovering the organisms in culture or the demonstration of antibodies to leptospires. Demonstration techniques have low sensitivity and specificity. Leptospires grow slowly and the positivity rate in culture is very low. Although microscopic agglutination test has been the cornerstone of serological diagnosis, the procedure is complex. New tests, like ELISA, dipstick test, lateral flow, etc, are relatively simple and rapid, but sensitivity is low during the early stages of the disease. The cross agglutination absorption test (CAAT) and typing with monoclonal antibodies (MCA) are the techniques used for serological characterization. These techniques are complicated and might not help in the case of certain serogroups. An alternate method for early diagnosis and characterization focuses on DNA-based techniques. Polymerase chain reaction (PCR), in situ hybridization etc are some of the methods used for early diagnosis, whereas restriction endonuclease analysis (REA), random amplified polymorphic DNA (RAPD) fingerprinting, arbitrarily primed PCR (AP-PCR), pulsed field gel electrophoresis (PFGE), ribotyping and DNA sequencing are useful for characterization. PCR is the most popular and quickest method for diagnosis. It can detect even if only a small number of organisms are present in a clinical sample. Fingerprinting tools such as RAPD, REA, RFLP, PFGE etc translate the complex genetic code into easily recognizable patterns, which facilitates characterization of the isolates up to sub-serovar level.
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PMID:Molecular tools in leptospirosis diagnosis and characterization of isolates. 1923 May 88